Mitotic control of RUNX2 phosphorylation by both CDK1/cyclin B kinase and PP1/PP2A phosphatase in osteoblastic cells

Arun Rajgopal, Daniel W. Young, Khawaja A. Mujeeb, Janet L. Stein, Jane B. Lian, Andre J. Van Wijnen, Gary S. Stein

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

Skeletal development and osteoblast maturation require the phenotype promoting activity of the transcription factor RUNX2, which controls both cell growth and differentiation in osteoblasts. We have recently shown that in actively proliferating cells RUNX2 regulates the expression of specific target genes as cells enter and exit mitosis. In this study, we addressed whether post-translational modifications of RUNX2 control its activity during mitotic exit. Western blot analysis of proteins from osteoblastic Saos-2 cells released from mitotic inhibition into early G1 show a phosphatase-sensitive shift in the mobility of RUNX2 in SDS gels. The slowly migrating hyper-phosphorylated form of RUNX2 is immunoreactive with a CDK related phospho-antibody (MPM2) only in mitotic cells and is converted into a faster migrating hypo-phosphorylated RUNX2 when cells complete mitosis. This conversion is inhibited by okadaic acid, an inhibitor of protein phosphatases 1 and 2 (PP1 and PP2A), but not by deltamethrin which blocks PP2B phosphatase. Mitotic phosphorylation of RUNX2 is sensitive to the CDK inhibitors roscovitine and olomoucine. Furthermore, RUNX2 can directly interact with CDK1 and is phosphorylated in vitro by the CDK1/cyclin B kinase complex. Hence, RUNX2 is hyper-phosphorylated by CDK1/cyclin B during mitosis, and dynamically converted into a hypo-phosphorylated form by PP1/PP2A-dependent dephosphorylation after mitosis to support the post-mitotic regulation of RUNX2 target genes.

Original languageEnglish (US)
Pages (from-to)1509-1517
Number of pages9
JournalJournal of cellular biochemistry
Volume100
Issue number6
DOIs
StatePublished - Apr 15 2007

Keywords

  • CDK1
  • Cyclin B
  • Mitosis
  • Okadaic acid
  • Osteoblast
  • Osteosarcoma
  • RUNX2
  • Roscovitine

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Mitotic control of RUNX2 phosphorylation by both CDK1/cyclin B kinase and PP1/PP2A phosphatase in osteoblastic cells'. Together they form a unique fingerprint.

  • Cite this

    Rajgopal, A., Young, D. W., Mujeeb, K. A., Stein, J. L., Lian, J. B., Van Wijnen, A. J., & Stein, G. S. (2007). Mitotic control of RUNX2 phosphorylation by both CDK1/cyclin B kinase and PP1/PP2A phosphatase in osteoblastic cells. Journal of cellular biochemistry, 100(6), 1509-1517. https://doi.org/10.1002/jcb.21137